PHOSPHATASES ALCALINES BACTERIENNES MODIFIEES ET LEURS APPLICATIONS.

摘要

Prodn. of a modified bacterial alkaline phosphatase (BAP) that has higher activity and similar thermal stability compared with the corresp. wild-type BAP comprises: (i) the preparation of an inactive chimeric alkaline phosphatase comprising the introduction into the sequence coding for the BAP, of at least two codons coding for two amino acid residues of the active site of a mammalian alkaline phosphatase; (ii) realisation of a random or directed mutagenesis on the gene coding for this inactive chimeric alkaline phosphatase; (iii) the expression of the alkaline phosphatases obtained in (ii); (iv) selection of bacterial clones expressing an alkaline phosphatase whose enzymatic activity is restored; and (v) the sequencing of the mutated alkaline phosphatases thus obtained and the selection of the mutation(s) of compatibility to be introduced into a wild-type bacterial alkaline phosphatase.